Prpf40b基因缺失诱发大鼠心脏结构异常

Prpf40b deletion causes cardiac structural abnormalities in rats

目的构建Prpf40b基因敲除大鼠,为该基因的生物学研究建立工具动物,同时初步探究该基因缺失后对心脏发育的影响。方法利用CRISPR/Cas9技术构建Prpf40b基因敲除大鼠,测序及PCR技术鉴定敲除大鼠构建成功及仔代大鼠基因型。通过超声影像技术分析敲除大鼠心脏结构形态和功能改变,随后通过病理组织学观察分析该基因敲除后对大鼠心肌显微形态的组织学影响。结果经PCR鉴定和测序比对,确认Prpf40b基因敲除大鼠构建成功。经超声影像学分析,与同窝阴性大鼠相比2月龄敲除大鼠心脏结构形态未见明显异常,而12月龄敲除大鼠收缩期及舒张期时心室腔内径及容积显著减小,同时射血分数显著减小。经病理组织学分析,与同窝阴性大鼠相比12月龄敲除大鼠心肌出现排列不齐,心肌纤维粗细不均及肌浆网扩张等现象。结论Prpf40b基因缺失可诱发大鼠心脏整体结构形态改变及心肌组织学异常。

Objective To establish Prpf40b knockout rats for biological research of the gene and explore the effect of gene deletion on development of the heart.Methods Prpf40b knockout rats were established by CRISPR/Cas9 technology.The genotypes of the founder rats and offspring were identified by sequencing and PCR.The cardiac structure and function of knockout rats were analyzed by ultrasound imaging technology.The microscopic morphology of the rat myocardium was analyzed by histopathological observation.Results PCR and sequencing confirmed successful establishment of Prpf40b gene knockout rats.Compared with the wt rats,ultrasound imaging analysis showed that the cardiac structure and morphology of 2-month-old knockout rats were not significantly abnormal;whereas the ventricular cavity diameter and volume of 12-month-old knockout rats decreased significantly in the systole and diastole,and the ejection fraction decreased significantly.Histopathological analysis showed that the myocardium of 12-month-old knockout rats was arranged irregularly,the thickness of myocardial fibers was uneven,and the sarcoplasmic reticulum was dilated.Conclusions Deletion of the Prpf40b gene induces changes in the overall structure and morphology of the rat heart and abnormal myocardial histology.